Effects of Phorbol Myristate Acetate on the Metabolism and Ultrastructure of Neutrophils in Chronic Granulomatous Disease

Abstract

A B S T R A C T Previous investigations have demonstrated that phorbol myristate acetate (PMA), the active principle of croton oil, stimulates alterations in normal polymorphonuclear leukocytes (PMN) that resemble closely the changes that develop in the cells after phagocytosis of bacteria. The present study has compared the effects of PMA and heat-killed bacteria on the oxygen uptake, glucose oxidation, nitroblue tetrazolium (NBT) reduction, and ultrastructure of normal neutrophils and PMN from six patients with chronic granulomatous disease (CGD). PMA stimulated oxygen consumption, hexose monophosphate shunt activity, and NBT reduction in normal cells but failed to produce similar effects in CGD neutrophils. However, PMA did induce formation of cytoplasmic vacuoles in the CGD cells similar to those observed in normal neutrophils. The results indicate that PMA is a useful nonparticulate agent for distinguishing between normal and CGD neutrophils and for studying basic mechanisms of phagocytosis in normal and abnormal PMN.

Stimulation of the oxidative metabolism of polymorphonuclear leucocytes by the calcium ionophore A23187

“…Recently increased cGMP concentrations have been demonstrated during PMNL phagocytosis [ 171. Phorbol myristate acetate increases cGMP in several cell types and has also been shown to stimulate the oxidative changes of phagocytosis in human leucocytes [3].…”

“…The increase in oxygen uptake and shunt activity can be detected within a few seconds after the addition of phagocytosable particles [ 11. In addition, several agents thought to interact with the PMN cell surface membrane, e.q. concanavalin A [2], phorbol myristate acetate [3], phospholipase C [4], endotoxin and detergents [5] provoke these oxidative changes. Thus we and others have proposed that contact between a phagocytic particle and the surface of the PMN may generate a chemical signal or signals which in turn mediate the characteristic metabolic accompaniments of phagocytosis [ 1,6], We have previously investigated the role of CAMP as a chemical signal.…”

Absence of cytochrome b reduction in stimulated neutrophils from both female and male patients with chronic granulomatous disease

“…The evidence for the involvement of this cytochrome b in the oxidase system is 2-fold. Stimulation of neutrophils with phorbol myristate acetate (PMA) activates the oxidase system [4] and results in the reduction and subsequent oxidation of this cytochrome b [5]. This cytochrome b was not found in reducedoxidised spectra of neutrophils from a group of patients with Chronic Granulomatous Disease (CGD) [6], a syndrome in which a predisposition to bacterial infection is associated with the absence of demonstrable oxidase activity in their neutrophils upon stimulation [7].…”

Differences in the association between the oxidase‐dependent activity and plasma membrane receptors for IgG, C3b and concanavalin A of human neutrophils

“…N~utraphil granulocytes fPMN) generate superoxide anions fctz) as part of an important defense system [l]e The generation of 0;: and other o~y~e~~deriv~ metabolites (OH', I+$&) are initiated at the surface tevef during p~~ti~i~-p~a~o~~~ interaction also in the absence of phagocytosis [2,3], A metabolic burst occurs also when PMN are exposed to soluble stimuli such as concanavalin A (ConA) [2], formylated peptides [4], CSa [5,6], phorbol myristate acetate (FMA) (7] and fatty acids [S], Htf~fiitfl PMN were isolated after dextran sedirne~tat~~~ as in [l I]_ After separation, washing and hypoto~ic lysis of contaminaiing er~~roc~e$s the PMN were suspended in Krebs-Ringer phosphate buffer containing 10 mM glucose (KRG), pH 7.2. Trypan blue exclusion was used to assay viability,…”